EP0327342B1 - Verfahren zur Herstellung von Erythritol - Google Patents

Verfahren zur Herstellung von Erythritol Download PDF

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Publication number
EP0327342B1
EP0327342B1 EP89300974A EP89300974A EP0327342B1 EP 0327342 B1 EP0327342 B1 EP 0327342B1 EP 89300974 A EP89300974 A EP 89300974A EP 89300974 A EP89300974 A EP 89300974A EP 0327342 B1 EP0327342 B1 EP 0327342B1
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EP
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Prior art keywords
erythritol
fermentation tank
culture broth
concentration
cells
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
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EP89300974A
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English (en)
French (fr)
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EP0327342A3 (en
EP0327342A2 (de
Inventor
Hiroyuki Horikita
Nobuo Hattori
Yahei Takagi
Gaku Kawaguchi
Toshihiro Maeda
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NGK Insulators Ltd
Mitsubishi Chemical Corp
Nikken Chemicals Co Ltd
Original Assignee
NGK Insulators Ltd
Mitsubishi Chemical Corp
Nikken Chemicals Co Ltd
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Publication of EP0327342A2 publication Critical patent/EP0327342A2/de
Publication of EP0327342A3 publication Critical patent/EP0327342A3/en
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Publication of EP0327342B1 publication Critical patent/EP0327342B1/de
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/04—Cell isolation or sorting
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/12—Purification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/813—Continuous fermentation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/818—Aeration or oxygen transfer technique
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/911—Microorganisms using fungi

Definitions

  • the present invention relates to a process for producing erythritol, which is a polyol, by using erythritol-producing microorganisms.
  • Erythritol is also called erythrit, and has the following characteristics:
  • Erythritol is a sugar alcohol, and demand for erythritol has recently been increasing because its sweetness is twice that of sucrose.
  • JP-A-60-110,295 and JP-A-61-31,091 disclose processes for producing erythritol from fermentable saccharides such as glucose with use of the erythritol-producing micro-organisms
  • US-A-3,756,917 discloses a process for producing erythritol from hydrocarbons.
  • EP-A-139592 describes method and apparatus for carrying out a batch-type fermentation for producing amino-acids. At a predetermined amino-acid concentration, the medium is filtered, and the filtration residue containing cells is recycled into the fermentation zone.
  • EP-A-243914 also describes a batch method and apparatus for fermentation of cell cultures.
  • the present invention has been accomplished to solve the above-mentioned problems, and to provide a process for producing erythritol in which production rate can be at a high level with a compact apparatus.
  • the production rate of erythritol can greatly be increased.
  • a substrate is continuously charged into a fermentation tank together with an inorganic salt such as KH2PO4, MgSO4, K2SO4, CaSO4, FeSO4, MnSO4, ZnSO4, (NH4)2HPO4, or CaCl2, a nitrogen source such as (NH4)2SO4, urea, NH4NO3, or NH4Cl, a nutrient source such as corn steep liquor, yeast extract, peptone, various amino acids, thiamine, or bitotin, while the liquid inside the fermentation tank is stirred by a stirrer 2 with blowing in of air clarified by an air filter 4 through an air feeding pipe 3 at the bottom.
  • an inorganic salt such as KH2PO4, MgSO4, K2SO4, CaSO4, FeSO4, MnSO4, ZnSO4, (NH4)2HPO4, or CaCl2
  • a nitrogen source such as (NH4)2SO4, urea, NH4NO3, or NH4Cl
  • a nutrient source such as corn steep liquor, yeast extract,
  • a cell separator 5 is shown arranged outside the fermentation tank.
  • the liquid inside the fermentation tank is separated into a concentrated liquid containing the erythritol-producing microorganisms and a clarified erythritol liquid containing no or a small amount of the erythritol-producing microorganisms by the cell separator 5.
  • the concentrated liquid is returned into the fermentation tank 1 through a line 6.
  • the clarified liquid is extracted outside the fermentation system as a fermented product.
  • the concentration of the cells in the liquid inside the fermentation tank gradually increases. Consequently, the production rate of erythritol reduces, and the cells are difficult to separate in the cell separator. Therefore, according to the present invention, a part of the liquid inside the fermentation tank is extracted outside through an extracting line 7 to control the amount of the cells in the tank.
  • the concentration of the cells is adjusted mainly by extracting a part of the culture broth outside the system by using the extracting pipe 7.
  • a part of the concentrated liquid is extracted outside through the above-mentioned line 6.
  • the thus extracted culture broth may be separated into a clarified liquid and a cell-concentrated liquid by the solid-liquid separator so that the clarified liquid may be utilized to obtain a fermented product.
  • the reason why the concentration of the dissolved oxygen in the culture broth inside the fermentation tank is limited to not less than 0.2 ppm is that the fermentation reaction is effected under aerobic conditions, and that, as shown in Fig. 3, since ethanol is produced in anaerobic fermentation if the concentration is less than 0.2 ppm, the production rate of erythritol lowers.
  • the graph shown in Fig. 3 was obtained by effecting experiments under the conditions that the concentration of the cells was kept at 100 g/l, while the concentration of oxygen dissolved was varied by changing the amount of air blown to various levels.
  • the cell separator use may be made of a liquid cyclone used in ordinary solid-liquid separation or a solid-liquid separator such as a precipitation separating tank. Noting the size of the erythritol-producing microorganisms, it is preferable to use a separated plate type (De Laval type) or inclined type (super decant type) centrifugal precipitation device which can continuously separate the cells from the liquid by utilizing centrifugal forces of 1,000 G to 10,000 G, a microfiltration membrane, or an ultra filtration membrane.
  • a separated plate type De Laval type
  • inclined type super decant type centrifugal precipitation device
  • a separating membrane of the membrane separator employed as the cell separator it is preferable to use a microfiltration membrane having pores of not more than 1 »m in diameter or an ultra filtration membrane having a cut-off molecular weight of not less than 10,000 for the purpose of assuredly separating the cells. If the pore diameter is more than 1 »m, the pores of the filter are likely to be plugged by the yeast, etc. Consequently, a permeation flux tends to lower in a short time. Similarly, if the fractionation molecular weight is not more than 10,000, the permeation flux drops. Thus, advantageous separation becomes impossible.
  • Fig. 4 show results obtained by testing characteristics of such separation membranes with use of a flat membrane type tester.
  • Reference numerals 1 through 4 denote filters having diameters of pores (3 » ⁇ 0.2 »)allotted thereto in Fig. 4, respectively, and reference numerals 5 through 9 show those having allotted a cut-off molecular weights (5,000 ⁇ 40,000), respectively.
  • the material of each of the filters is as follows: 1, 4 metal 2, 3 ceramic 5, 6, 7, 9 polyacryl nitrile 8 polyester sulfone In the tests, a liquid containing 94 g/l of cells, 19% of erythritol and 2% of glucose was continuously passed under pressure of 1 kg/cm2 for 2 hours.
  • the liquid inside the fermentation tank can be separated into the cell-containing concentrated liquid and the filtrate containing erythritol by using an appropriate separation membrane 5.
  • the amount of the liquid extracted from the fermentation tank through the line 7 is determined such that the concentration of the cells in the fermentation tank is kept at 40 to 200 g/l when calculated as a dried cell weight.
  • the amount of the liquid extracted is set at not greater than that of the clarified liquid to be extracted. As shown in Fig. 5, if the amount of the cells is less than 40 g/l, the number of cells becomes insufficient, reducing the production rate of erythritol. On the other hand, if the amount is more than 200 g/l, the number of cells becomes excessive, also reducing the production rate of erythritol while requiring a greater amount of oxygen, and reducing the production rate per unit power. For this reason, the amount of the cells is set preferably at 40 to 200 g/l, more preferably 80 to 150 g/l.
  • Fig. 2 shows a fermentation system to be used in the second aspect of the present invention.
  • a separating membrane 5 is of tubular type, and is arranged inside the fermentation tank 1.
  • the line 6 in Fig. 1 can be omitted.
  • a filtering pressure may be applied to the filtering membrane 5 by sealing an upper face of the fermentation tank 1 and pressurizing the liquid surface.
  • Examples 1 and 2 belong to the first aspect of the present invention, and Example 3 to the second aspect of the invention.
  • Erythritol-producing microorganisms (Aureobasidium sp. SN-G42 which is publicly available, deposit no. FERM BP-1430) were cultivated in 200 ml of a liquid culture medium containing 30% (w/v) of glucose and 1% (w/v) of yeast extract at 35°C for 72 hours with shaking, which was added to 3.8 l of an initial culture medium containing 40% (w/v) of glucose and 2% (w/v) of yeast extract. The mixture was batch cultivated at 35°C and a stirring rate of 800 rpm while air was being passed at 1 vvm.
  • 1.5 l of seed cultures of erythritol-producing microorganisms (Aureobasidium sp. SN-G42), which had been cultivated in a culture broth containing 30% (w/v) of glucose and 0.675% (w/v) of yeast extract in a rotary shaker at 30°C for 72 hours with shaking, were added to 25 l of an initial culture medium which was placed in a 50 l fermentation tank and which contained 40% (w/v) of glucose and 6.7% (w/v) of corn steep liquor and was adjusted to pH 4.2.
  • the mixture was cultivated under sufficiently aerobic conditions of 35°C, forced stirring at 600 rpm and pressure of 0.5 g/cm2, while aseptic air was fed at a rate of 37.5 l/min.
  • the feed rate to the centrifugal precipitation device was dropped to 0.96 l/h, and at the same time the amount of the clarified liquid extracted from the centrifugal separator was reduced to 0.32 l/h.
  • the culture broth inside the fermentation tank was directly extracted at 0.26 l/h. The fermentation system reached a steady state 250 hours after the cultivation, and the cultivation was further continued for 150 hours.
  • the average concentration of erythritol in a mixed liquid consisting of 0.32 l/h of the clarified liquid and 0.26 l/h of the extracted culture broth was 194 g/l, and the yield and the production rate of erythritol were 48.5% and 4.5 g/l/h, respectively.
  • Erythritol-producing microorganisms (Aureobasidium sp. SN-G42) were cultivated in 200 ml of a liquid culture medium placed in a 500 ml conical flask and containing 30% (w/v) of glucose and 1% (w/v) of yeast extract at 35°C for 48 hours with shaking, which was added to a 7 vol l fermentation tank.
  • the fermentation tank contained 3 l of a culture medium containing 40% (w/v) of glucose and 8% (w/v) of corn steep liquor, and was equipped with a cell separator (denoted by a reference numeral 5 in Fig. 2).
  • batch cultivation was effected under conditions of 35°C, pH 4.2 and a stirring rate of 1,000 rpm, while air was passed at 1 vvm.
  • concentration of glucose in the culture medium reached 5% or less after 70 hours
  • a culture medium containing 40% (w/v) of glucose and 8% (w/v) of corn steep liquor was fed at 83.3 ml/h as a substrate.
  • a cultivation filtrate was also extracted using a filter at the same rate, continuous cultivation was effected.
  • the extracting rate of the cultivation filtrate was dropped to 41.65 ml/h and at the same time the culture broth was extracted directly from the fermentation tank at a rare of 41.65 ml/h so as to keep the concentration of the cells constant. Then, cultivation was continuously effected for 167 hours, while the concentration of the cells was kept at 100 g/l. During the cultivation, the average production rate and the yield of erythritol were 5.1 g/l/h and 50%, respectively.
  • the present invention has succeeded in raising the production rate of erythritol to for example twice the conventional rate while maintaining the concentration of the cells at an appropriate level, by controlling the concentration of dissolved oxygen, separating the cells, and extracting the culture broth inside the fermentation tank to the outside.
  • the entire fermentation apparatus can be made compact with no need to use a large scale of a fermentation tank as in the conventional batch type production.

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Claims (3)

  1. Verfahren zur kontinuierlichen Herstellung von Erythrit durch Kultivieren Erythrit erzeugender Mikroorganismen in einer Kulturbrühe unter aeroben Bedingungen in einen Erythrit erzeugenden System, wobei das Verfahren folgende Schritte umfaßt:
    das Halten der Konzentration an in der Kulturbrühe innerhalb eines Fermentationstanks gelöstem Sauerstoff auf nicht unter 0,2 ppm;
    das Auftrennen eines Teils der Kulturbrühe in eine konzentrierte Flüssigkeit, in der die Konzentration an Zellen erhöht ist, und eine geklärte Flüssigkeit, mittels eines Zellseperators;
    das Rückführen der konzentrierten Flüssigkeit zum Fermentationstank;
    das Regulieren einer Menge der aus dem Erythrit erzeugenden System extrahierten geklärten Flüssigkeit und einer Menge der Kulturbrühe und/oder der aus dem Erythrit erzeugenden System extrahierten konzentrierten Flüssigkeit, sodaß die Konzentration der Zellen in der Kulturbrühe im Fermentationstank in einem Bereich von 40 bis 200 g/l, berechnet als Gewicht getrockneter Zellen, gehalten wird;
    und das Gewinnen von Erythrit aus der geklärten Flüssigkeit.
  2. Verfahren zur kontinuierlichen Herstellung von Erythrit durch Kultivieren von Erythrit erzeugenden Mikroorganismen in einer Kulturbrühe unter aeroben Bedingungen, wobei das Verfahren folgende Schritte umfaßt:
    das Halten der Konzentration an in der Kulturbrühe in einem Fermentationstank gelöstem Sauerstoff auf nicht weniger als 0,2 ppm;
    das Abtrennen einer Erythrit enthaltenden geklärten Flüssigkeit von der Kutlurbrühe durch einen innerhalb des Fermentationstanks angeordneten Zelltrenner;
    das Regulieren einer Menge der aus der Kulturbrühe extrahierten geklärten Flüssigkeit und einer Menge der aus dem Fermentationsbehälter extrahierten Kulturbrühe, sodaß die Konzentration der Zellen in der Kulturbrühe im Fermentationstank in einem Bereich von 40 bis 200 g/l, berechnet als Gewicht getrockneter Zellen, liegt;
    und Gewinnen von Erythrit aus der geklärten Flüssigkeit.
  3. Verfahren nach Anspruch 1 oder 2, worin die Erythrit erzeugenden Mikroorganismen Aureobasidium, sp. SN-G42 (FERM BP-1430) sind.
EP89300974A 1988-02-03 1989-02-01 Verfahren zur Herstellung von Erythritol Expired - Lifetime EP0327342B1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP24813/88 1988-02-03
JP63024813A JPH0734749B2 (ja) 1988-02-03 1988-02-03 エリスリトールの製造方法

Publications (3)

Publication Number Publication Date
EP0327342A2 EP0327342A2 (de) 1989-08-09
EP0327342A3 EP0327342A3 (en) 1990-08-22
EP0327342B1 true EP0327342B1 (de) 1995-07-19

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EP89300974A Expired - Lifetime EP0327342B1 (de) 1988-02-03 1989-02-01 Verfahren zur Herstellung von Erythritol

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US (1) US4923812A (de)
EP (1) EP0327342B1 (de)
JP (1) JPH0734749B2 (de)
DE (1) DE68923464T2 (de)

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DE69720379T2 (de) * 1996-12-02 2004-02-12 Mitsubishi Chemical Corp. Verfahren zur Herstellung von Erythritol
KR100541578B1 (ko) * 1997-12-04 2006-04-06 미쓰비시 가가꾸 가부시키가이샤 에리트리톨 생산방법
KR100246820B1 (ko) * 1997-12-30 2000-03-15 정수련 신균주 트리고높시스 배리아빌리스에 의한 에리쓰리톨의 제조방법
BR9909584A (pt) * 1998-04-14 2000-12-19 Dsm Nv Filtração por membrana
FR2780414B1 (fr) * 1998-06-24 2001-06-08 Roquette Freres Procede de production d'erythritol par fermentation discontinue alimentee repetee
KR100271137B1 (ko) * 1998-06-24 2000-11-01 유병택 신규 미생물 트리코스포로노이데스 마디다 디에스911 및 이 균주를 이용한 에리스리톨의 제조방법
AU1234400A (en) * 1998-10-27 2000-05-15 Cargill Incorporated Method for purifying a polyol product stream
FR2793498B1 (fr) * 1999-05-11 2001-07-27 Roquette Freres Procede de production d'arabitol par fermentation continue
US6300107B1 (en) * 2000-06-02 2001-10-09 Food Industry Research & Development Institute Erythritol-producing yeast strains
CN1335391A (zh) * 2000-07-21 2002-02-13 食品工业发展研究所 生产赤藓糖醇的酵母菌株
JP5098122B2 (ja) * 2001-04-27 2012-12-12 三菱化学株式会社 連続培養によるエリスリトールの製造方法
US7704248B2 (en) * 2005-12-21 2010-04-27 Boston Scientific Scimed, Inc. Ablation device with compression balloon
AT504230B1 (de) 2006-10-03 2008-06-15 Jungbunzlauer Austria Ag Verfahren zur herstellung von erythrit
JP5287029B2 (ja) * 2007-08-22 2013-09-11 東レ株式会社 連続発酵による化学品の製造方法
JP2009296921A (ja) * 2008-06-12 2009-12-24 Toray Ind Inc 連続培養装置および化学品の製造方法
JP5659466B2 (ja) * 2009-08-07 2015-01-28 東レ株式会社 連続培養による化学品の製造方法および製造装置
JP2011092041A (ja) * 2009-10-28 2011-05-12 Kansai Chemical Engineering Co Ltd エタノール生産微生物の連続培養発酵装置
US8801682B2 (en) 2010-01-27 2014-08-12 Human Med Ag Apparatus for separating tissue cells from a fluid
DE102010001292B4 (de) * 2010-01-27 2013-08-01 Human Med Ag Vorrichtung zum Trennen von Gewebezellen aus einer Flüssigkeit
EP2436772A1 (de) * 2010-09-30 2012-04-04 Annikki GmbH Verfahren zur Herstellung von Erythritol
JP7154093B2 (ja) * 2018-10-04 2022-10-17 佐竹マルチミクス株式会社 培地抜出機構を有する培養装置及びこの培養装置の培地の交換方法

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Also Published As

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EP0327342A3 (en) 1990-08-22
DE68923464D1 (de) 1995-08-24
DE68923464T2 (de) 1996-03-21
JPH0734749B2 (ja) 1995-04-19
JPH01199584A (ja) 1989-08-10
US4923812A (en) 1990-05-08
EP0327342A2 (de) 1989-08-09

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